Novel 15-dehydro-halo-methyl-progesterones



United States Patent 3,183,150 NOVEL 15-DEHYDRO-HALO-METHYL- PROGESTERONES Fritz von Werder, Klaus Briickner, Kari-Heinz Bork, and

Harald Metz, Darmstadt, Germany, assignors to E. Merck Aktiengesellschaft, Darmstadt, Germany No Drawing. Filed Sept. 10, 1962, Ser. No. 222,666 Claims priority, application Germany, Sept. 16, 1961,

M 50,326; Sept. 28, 1961, M 50,415, M 50,416,

6 Claims. (Cl. 167-53) The present invention relates to new and useful steroids of the pregnane series.

It is an object of this invention to provide novel compounds having a valuable physiological activity as well as compositions containing these compounds together With pharmaceutically acceptable excipients.

It is a further object of this invention to provide a ne process for producing said compounds.

The new compounds of this invention includes those represented by the Formula I CHgRl do I , orn nj i m-on x (I V wherein R R R X and Y have the above meaning. Likewise, the A A or A -derivatives thereof may be used.

The starting com-pounds of Formula II are readily available from the corresponding 16-methylene-l7a-hydroxy steroids some of which are described in Tetrahedron Letters, No. 16, 1960, pp. 2l-32, or which may be obtained from the compounds described in South African Patent 264/ 61 by saponification of the 17a-ace- 3,183,150 Patented May 11, 1955 toxy group and, if desired, by substituting a hydrogen atom in the 21-position by a hydroxyl or acyloxy group according to standard methods. By treatment with N- chloroor N-bromo-succinimide, such 16-methylene-17ahydroxy compounds are converted into the 16-bromoor 16 chloromethyl-l6u,l7 x-o ddo-steroids according to Formula 11. Starting with the 16-bromomethyl compounds, the 16-fluoromethyl-l6a,l7u-oxido steroids of Formula II are prepared by treatment with silver fluoride in acetonitrile or with potassium fluoride in diethyleneglycol.

According to the new process of this invention, a compound of Formula II is treated with a strong inorganic acid in the presence of water and an inert organic solvent miscible with water to form the corresponding 15-dehydrol6-halomethyl-l7a-hydroxy steroid of Formula III ---OH ]-011,X /g

(III) or the A A or A -derivative thereof wherein R R R X and Y have the above significance.

Inorganic acids suitable for the process of the present invention are, for example, perchloric acid, hydrogen halides, i.e. hydrochloric, hydrobromic, hydroiodic and hydrofluoric acid, sulfuric acid etc.

The presence of water during the splitting of the oxido ring seems to be necessary for the course of the reaction. It is preferred that the mol ratio of water to organic solvent is approximately from about 3:1 to 1:50.

Suitable inert organic solvents for this reaction are, for example, tetrahydrofuran, acetone, methanol, ethanol, acetonitrile, dimet-hylforma-mide, dioxane, or glacial acetic acid. The preferred range of the reaction temperature is from 0 C. to the boiling point of the used solvent. The reaction times vary from /2 to 24 hours depending on starting material and temperature. Generally, the reaction mixture is allowed to stand for about /2 to 2 hours at room temperature. Then it is poured into water whereby the desired product is precipitated. It may be purified by recrystallization or by chromatographic methods.

To prepare the new compounds of this invention, the splitting of the 16-halomethyl-l6a,l7a-oxido steroids in the presence of the acids and solvents as indicated above is the main reaction and may be carried out at any desired reaction stage. Thus, it may be the final step in the preparation of the useful compounds of Formula I, or, likewise, an intermediate step which is followed by one or more further reactions well known in steroid chemistry.

For example, a hydroxyl group may be introduced into the ll-position of a 15-dehydro-16-halomethyl-17ahydroxy steroid of the Formula III wherein R and Y are hydrogen by usual microbiological methods. All microorganisms suitable for such reactions may be employed, for example, fungi of the genus Curvularia, Mucor, Stachylidium and Streptomyces (introducing a hydroxyl group at the llfl-position) or fungi of the genus Absidia, Cunninghamella, Fusarium, Mucor, Pencillium, Rhizopus (introducing a hydroxyl group at the Ila-position). The hydroxylation is effected according to stand- $9 ard procedures. The starting material is added to a culture of the microorganism which grows in a suitable nutrient solution at optiumum temperature and with aeration. After to 48 hours, the ll-hydroxy steroid is isolated from the reaction mixture, preferably by extraction with a suitable organic solvent such as chloroform or methylene chloride.

The obtained llhydroxylated compounds may subsequently be oxidized to from the corresponding ll-keto steroids. For this oxidation, mild oxidizing agents are preferred. For example, chromic acid anhydride in glacial acetic acid or a mixture of chromic acid anhydride and pyridine or a mixture of sodium bichromate, sulfuric acid and acetone may be used. The ll-keto steroids are isolated from thereaction mixture by extraction or by precipitation with water.

Furthermore, a double bond may be introduced at any reaction stage into the 1,2-position. The l,2-dehy drogenation can be effected chemically or microbiologically. As chemical dehydrogenation agents, there are particularly suitable 2,3-dichloro-5,6-dicyano-benzoquinone or selenium dioxide.

When using selenium dioxide, an alcohol such as t-butanol, t-amyl alcohol, or ethyl acetate is employed as solvent. The reaction can be accelerated by the addition of small amounts of glacial acetic acid. Advantageously, the reaction mixture is refluxed for about 12 to 48 hours. The precipitated selenium is separated.

For a dehydrogenation with 2,3-dichloro-5,6-dicyano benzoquinone, benzene and dioxane, are particularly suitable as solvents.

For the microbiological 1,2-dehydrogenation, there can be used all the microorganisms customary for this purpose. Bacillus sphaericus var. fusiformis, Corynebacterium simplex and Fusarium solani are particularly suitable.

For the dehydrogenation, the starting material is added to a submerged culture of the microorganism employed which grows in a suitable nutrient solution at optimum temperature and with strong aeration in accordance with the customary methods of the fermentation art. Instead of growing cultures, suspensions of the microorganisms in buffer solutions can also be used, the method being otherwise the same. The course of the reaction is observed chromatographically and the fermentatioin solution extracted, for instance with chloroform, after complete reaction of the starting material.

In some cases, 2l-O-acyl compounds are converted into 21-OH compounds in the course of a microbiological process as described above.

To introduce a double bond into the 6,1-position, a compound of Formula I or III which is saturated in the 6-position is reacted with chloranil. One advisedly operates in a solvent having a boiling point of about 30 to 150 C. As solvents, the following are, for instance, suitable: alcohols, such as ethanol, tert.-butanol or tert.- amyl alcohol, methyl acetate, ethyl acetate, dioxane, glacial acetic acid, benzene, tetrahydrofuran, acetone, etc.

The compounds of the Formula III wherein R is hydrogen can be converted at any desired reaction step into the corresponding 21-hydroxyl or 21-acyloxy compounds. According to the most common method known in the art, the starting material is treated subsequently with an alkaline solution of iodine and with an alkali metal acylate, preferably potassium acetate. The 2l-acyloxy compound thus obtained may be saponified, if desired, to form the corresponding 2l-alcohol. The saponification is effected in the usual manner, for example, by an aqueous solution of sodium bicarbonate.

To prepare the 9a-fluoro-compounds of the invention, the usual sequence of reactions may be employed. For example, a compound of the Formula III wherein R3 and Y are hydrogen can be hydroxylated microbiological- 1y at the ll-position. Upon treatment with a dehydrating agent, the corresponding 9,11-dehydro compound is obtained which is converted into the 95,11B-oxido compound with standard procedures. Reaction with hydrogen fiuoride results in the formation of the corresponding 9wfiuoro-llfi-hydroxy compound.

The 2l-position hydroxyl group present in any of the molecules whether starting material, intermediate or product may be readily esterified by standard methods. All acids or their derivatives are suitable which form physiologically acceptable esters. Among the large number of acids to be used, the following are named by way of illustration: acetic acid, propionic acid, butyric acid, trimethyl acetic acid, t.-butyl acetic acid, cyclopentylpropionic acid, phenylpropionic and phenylacetic acid, capronic acid, caprylic acid, palmitic acid, undecylenic acid, benzoic acid, chloroacetic acid, diethylaminoacetic acid, aspartic acid, oxalic acid, succinic acid, phosphoric acid, sulfuric acid. In the event that the esterified 21-hydroxyl group is one derived from a dibasic acid, it is often advantageous to treat such esters with an alkali metal or alkaline earth metal hydroxide to prepare the corresponding salt. Such salts are especially useful because of their increased solubility in water.

To obtain compounds of the Formula I wherein R is an acyl group, the corresponding Not-hydroxyl derivatives are acylated in accordance with known methods customary for tertiary hydroxyl groups. Preferably, a mixture of acetic anhydride and glacial acetic acid, including an addition of p-toluenesulfonic acid, is employed as acylating agent. For 17a-acyloxy derivatives of carboxylic acids of higher carbon content, such as propionic or capronic acid, the corresponding acids and acid anhydrides are used.

The products and methods in accordance with the present invention make it possible to provide the physiologically active valuable 15-dehydro-l6-halomethyl compounds of Formula I. The new compounds can be used in veterinary and human medicine as therapeutic agents. The derivatives of the corticoid series possess an increased antiphlogistic activity which is tested according to the method described by R. Hotovy et al. in Archives Internationales de Pharmacodynamie et de Thrapie, vol. 111, p. 420 (1957); The new compounds of this invention belonging to the progesterone series exhibit an increased gestagenic activity. Some of them, particularly those substituted by an 17u-acyloxy-group, are characterized by extremely good oral effectiveness. In the Clauberg test on rats (cf. Journal of Physiology, vol. 83, p. (1934) the new compounds were compared to known gestagenes.

The active compounds of this invention may be administered alone or in combination -with acceptable pharmaceutical carriers, the choice of which is determined by the desired route of administration. They can be worked up into all forms of application which are customary for pharmaceutical purposes, such as pills, tablets, dragees, suppositories, emulsions, suspensions or injection solutions. Usual adjuvants, fillers, solvents or solubilizers may be added if desired. In general, the pharmaceutical preparations contain from 0.1 to 70 mg. of the active ingredient. Thus, tablet-s usually contain about 0.4 to 6 mg. in dosage unit form whereas ampoules contain a somewhat higher dose, such as from 4 to 60 mg.

The dosage of the compounds of the corticoid series is of approximately the same order as the dosage of prednisolone, and the compounds are useful to treat all types of pathological conditions often treated with prednisolone. Because of their great activity, the doses are sometimes even lower than those of prednisolone. As to the compounds of the progesterone type, these ones may be used for all indications of 17a-acetoxy progesterone or hydroxy-6a-methyl-progesterone. For instance, the substances are suitable as means for combating threatening abortion, for restoring the uterus mucous membranes or in functional uterine bleeding.

The following examples are given solely for illustration and are not to be construed as limitations of this invention. As will be recognized by those skilled in the art, there are many modifications of the processes for the preparation of the new compounds which are intended to be within the full spirit and scope of this invention. Especially, the sequence of the reaction steps may be varied largely, depending on the used starting materials and the desired product.

Example 1 Example 2 5 g. of 16B-fluoromethyl-16a,17a-oxid0-4-pregnene- 3,20-dione-1 1t3,21-diol are allowed to stand for 2 hours at room temperature with 75 ml. of dioxane and 25 ml. of concentrated hydrochloric acid (37%). The reaction mixture is poured into water. The precipitate is separated and chromatographed through 250 ml. of magnesium silicate. The fractions proved to be uniform by thin layer chromatography are combined and evaporated under jreduced pressure. The residue is the 16-fiuoromethyl-4,15-pregnadiene-3,20-dione 11,8,17a,21 triol. M.P. 209-211 (3.; [04] +69.7 (dioxaue);

Am. 240 m rrg 440 Example 3 According to the method described in Example 2, the 16/3-fluoromethyl-16a,17a-oxido-l,4 pregnadiene 3,20- dione-llfi,21-diol-21-acetate is split to form 16-fluoromethyl-1,4,15-pregnatriene 3,20-diOI16-11B,l7a,21-l21'i0l- 21-acetate. M.P. 208-209" C.; [orb +47.3 (dioxane);

A 242 m E1? 382 Example 4 (a) In accordance with the method described in Example 1, the lfl-fluoromethyl-l6a,l7a-oxido-4-pregnene- 3,20-di0ne is split off to form 16-fiuoromethyl-4,l5-pregnadiene-l7a-ol-3,20-dione. M.P. 219-220 C.; [ab 8 (chloroform); x 240 m 6:1765Q.

The obtained product is acetylated as described in Example 18(b) to form 16-fluoromethyl-4,15-pregnadiene-17a-ol-3,20 dione-17-acetate.

(b) 2 g. of 16-fluorornethyl-4,l5-pregnadiene-l7a-ol- ,3,20-dione-l7-acetate are dissolved in ml. of tetrahydrofuran and refluxed for 13 hours with 1.6 g. of chloranil. The reaction mixture is diluted with water and extracted with chloroform. The combined chloroform extracts are washed subsequently with sodium hydroxide (2 N) and with water. Upon evaporation, the 16-fluoromethyl-4,6,15-pregnatriene-17ot ol 3,20-dione-17-acetate is obtained.

Example 5 44 g. of l-chloromethyl-16a,17a-oxido-progesterone are dissolved in 1.76 l. of dioxane. Upon addition of 880 ml. of hydrochloric acid (37%), the mixture is allowed to stand for half an hour at room temperature. Then it is poured with stirring into 18 l. of Water. The precipitate is filtered off, Washed with water, dried and dissolved in ml. of a mixtureof benzene and chloro form (1:1). The solution is chromatographed through 1.41 kg. of silica gel. The fractions proved to be uniform by thin layer chromatography are combined and the solvents are evaporated. The residue of 16-chlororefluxed for 3 hours.

f5 methyl-4,1S-pregnadiene-17a-ol-3,20-dione is recrystallized from ether. M.P. l98-l99 C.; 15 (chloroform);

h 239-240 mp, Ei'f' 480 Example 6 3 g. of 6a-methyl-16a,17a-oxido-l6-fiuorornethyl-4- pregnene-3,20-dione are dissolved in 45 ml. of dioxane. Upon addition of 7.5 ml. of perchloric acid (70%) and 7.5 ml. of Water, the solution is allowed to stand for 2 hours at room temperature and then is worked up as described in Example 5 whereby the 6u-methyl-16-fluoromethyl-4,15-pregnadiene17o:-ol-3,20-dione is obtained.

Example 7 (a) According to the method of Example 1, the 16- chloromethyl-4,1S-pregnadiened7a-ol-3,20-dione is obtained from 16-chloromethyl-16p,17a-oxido-4-pregnene- 3,20-dione. Ml. 198-199 C.; M1 15 (chloroform);

A 239-240 m E'Ff 480 (b) To a suspension of 6.5 g. of 16-chloromethyl-4,15- pregnadiene-17or-ol-3,20-dione in 98 ml. of tetrahydro- 'furan and 59 ml. of methanol, 9.8 g. of iodine and 98 g.

of CaO are added in small portions within 3 hours. The reaction mixture is poured into 2 l. of ice water containing 32 ml. of glacial acetic acid. The precipitate is filtered off, dried and dissolved in 550 ml. of acetone. The solution is refluxed for 20 hours with 33 g. of potassium acetate. The acetone is partially evaporated under reduced pressure. Upon addition of water, the precipitate is filtered off and refluxed for 2 hours with a mixture of ml. of methanol, 3.25 g. of sodium pyrosulfite and 48.5 ml. of water. The reaction mixture is cooled and diluted with water. The precipitated 16-chloromethyl-4,15 pregnadiene l7u,21 did-3,20- dione-21-acetate is filtered 01f, dried and recrystallized from methanol.

(c) 12 g. of 16-chloromethyl-4,15-pregnadiene-17ot,21- diol-3,20-dione-2l-acetate' are dissolved in 960 ml. of methanol. Upon addition of 480 ml. of an aqueous potassium bicarbonate solution (5%), the mixture is After cooling, crystals of 16- chloromethyl-4, l S-pregnadiene-17a,2l-diol-3 ,20-dione are obtained which are recrystallized from acetone hm. 239-240 m Er? 460 (ethanol) li -l (d) In a fermentation vessel 15 l. of a nutrient solution containing 5% glucose, 0.1% yeast extract, 0.05% soybean meal, 0.3% NaNO 0.05% MgSO -7H O, 0.1% KH PO 0.05% KCl, 0.001% FeSO -7H O are'inoculated with 750 ml. of a culture of Fusarium sp. -The culture grows with vigorous stirring and aeration at 28 C. After 24 hours, 5 g. of 16-chloromethyl-4,15-pregnadiene-l7a,21-diol-3,20-dione in 40 ml. dimethylformamide are added. As soon as the paper chromatograrn no longer shows any starting material, the culture is extracted three times with 101. of chloroform. The chloroform extracts are evaporated; the residue is washed with petroleum ether and the 1S-dehydro-l6-chloromethyl-llepic-hydrocortisone recrystallized from acetone.

A 241 m E1? 430 (e) 2 g. of 1S-dehydro-16-chloromethyl-1l-epi-hydrocortisone are dissolved in 10 ml. of pyridine and 0.36 g. of glacial acetic acid anhydride. After standing for 15 hours at room temperature, the solution is poured into Water and extracted three times with chloroform. The

chloroform extracts are neutralized with a solution of x 238-239 mp, E1 13 365 (g) 1 g. of 15-dehydro-16-chloromethyl-cortisone-Z1- acetate is refluxed with 25 ml. of methanol. To the boili'ng solution, a hot solution of 0.23 g. of sodium bicarbonate in 5 ml. of water is added. After boiling for 7 minutes, the solution is poured into 300 ml. of water and the precipitated crude product is filtered under suction. Upon recrystallization from acetone the pure 15-dehydro- 16-chloromethyl-cortisone is obtained.

A 238 mp, El l'm. 375

(h) In a fermentation vessel, 15 l. of a nutrient solution containing of 0.1% yeast extract, pH 6.8, are inoculated with 1.5 l. of a culture of Corynebacterium simplex. The culture is grown with constant stirring and aeration at 28 C. After 48 hours, 7.5 g. of 15-dehydro- 16-chloromethyl-cortisone in 300 ml. of methanol are added. The dehydrogenation is controlled by paper chromatography and is usually finished after 10-14 hours. The solution is extracted three times with chloroform; the extracts are evaporated and the -dehydro-16-chloromethylp'rednisone is recrystallized from acetone.

m. 239 m llis]. 0

Example 8 (a) 16-chlorornethyl-4,15-pregnadiene-17a,21-diol-3,20- dione-21-acetate are prepared according to Example 1 from 16fi-chl0romethyl-16a,l7a-oxide-4-pregnene-21-ol-3, -dione-21-acetate.

(b) According to the method described in Example 7(a), the 16-chloromethyl-4,15-pregnadiene-17u,21-diol-3 20-dione-21-acetate is saponified to form 16-chloromethyl- 4,15-pregnadiene-17a,21-diol-3,20-dione.

(c) In a fermentation vessel, 15 l. of a nutrient solution containing 5% malt extract, 1% saccharose, 0.2% NaNQ 0.1% K HPO 0.05% MgSO 0.05% KCl and 0.005% FeSO pH 7.0, are inoculated with 800 ml. of a culture of Curvularia lu'nata (Warker) Boadijn. After growth for 24 hours at 28 C., 5 g. of lfi-chloromethyl- 4,IS-pregnadiene-17a,21-diol-3,20-dione in 40 ml. dimeth ylformamide are added. As soon as the paper chromatogram no longer shows any starting material, the culture is extracted three times with chloroform. The chloroform extracts are evaporated and the residue is chromatographed through silica gel. The eluate running ofi with chloroform/ethyl acetate (1:3) contains 15-dehydro-16- chloromethyl-hydrocortisone.

(d) 5 g. of 15-dehydro-16-chloromethyl-hydrocortisone are heated with 30 ml. of pyridine and 30 ml. of acetic acid anhydride for 1 hour on the steam bath. The solution is poured into water and the precipitated 15-dehydro- 16-chloromethyl-hydrocortisone-2l-acetate is filtered off and recrystallized from acetone.

A 241 mp, E1 5 395 (e 3.5 g. of IS-dehydro-16-chloromethyl-hydrocortisone-21-acetate and 3.5 g. of 2,3-dichloro-5,6-dicyanobenzoquinone are dissolved in 70 ml. of dioxane and refiuxed for 6 hours. The mixture is diluted with chloroform and washed subsequently with 30 ml. of aqueous sodium hydroxide (1 N) and water. The solution is dried and evaporated. The 1S-dehydro-16-chloromethyla C9 prednisolone-Zl-acetate is recrystallized from acetone/ ether.

Mm. 242 u, 3

(e In a fermentation vessel 15 l. of a nutrient solution containing 1% yeast extract, pH 6.8, are inoculated with 0.5 l. of a culture of Bacillus sphaericus. The culture is grown at 28 C. and after 10 hours 7.5 g. of 15- dehydro-l6 chloromeythl-hydrocortisone in 300 ml. of methanol are added. After 28-36 hours dehydrogenation is complete. The reaction mixture is treated according to Example 7(h). The 15-dehydro-16 chloromethyl-prednisolone is recrystallized from acetone.

Example 9 (a) According to the method described in Examples 7(a) and (b), the 16-fluoromethyl-4,15-pregnadiene-17a, 21-diol-3,20-dione-2l-acetate is prepared from 16-fluoromethyl-16a,17a-oxido-4-pregnene-3,20-dione.

r 239, 5 mp, Ei't... 395

(b) 12 g. of 16-fiuoromethyl-4,15-pregnadiene-17a21- diol-3,20-dione-2l-acetate and 480 ml. of potassium bicarbonate (aqueous solution, 5%) are dissolved in 960 ml. of methanol and refluxed for 3 hours. Upon cooling,

the 16-fluoromethyl-4,15-pregnadiene-17a,21-diol 3,20-

dione precipitates. It is recrystallized from acetone.

m. 240 u, lt... 435

(1c) According to the method described in Example 8 (c) the 16-fluoromethyl-1S-dehydro-hydrocortisone is obtained from 16-fluoromethyl-4,15-pregnadiene-17a,21-diol 3,20- dione. M.P. 210211 C.; [@1 9 +69.7 (dioxane);

(d) 5 g. of 16-fiuor0methyl-1S-dehydro-hydrocortisone are heated with 30 ml. of pyridine and ml. of acetic anhydride for 1 hour on the steam bath. The solution is poured into Water, and the precipitated 16-fluoromethyl- 15-dehydro-hydrocortisone-2l-acetate is filtered oil and recrystallized from acetone.

X 241 m Ei' g 418 (e,) According to the method described in Example 8(e the lS-dehydro-l6-fluoromethyl-prednisolone is prepared from lS-dehydro-16-fiuoromethyl-hydrocortisone. l /LP. 224225 C., [OL]D+22.8 (dioxane);

It is converted by usual methods into the corresponding 21-tert.-butylacetate.

X 242 mp, E1 5 332 (0 According to the method described in Example 8 (e the 16-fluoromethyl-1S-dehydro-prednisolone-Zlacetate is prepared from 16-fluoromethyl-1S-dehydro-hydrocortisone-Zl-acetate. M.P. 208-209 C.;

(f) 2.4 g. of 16-fiuoromethyl-IS-dehydro-prednisolone- Zl-acetate are dissolved in 24 ml. of pyridine and added to a solution of 2.4 g. of chromic acid anhydride in 24 m1. of pyridine. The mixture is allowed to stand overnight and worked up in the usual way whereby the 16-fluoromethyl-15-dehydro-prednisone2l-acetate is obtained.

Upon saponification, the 16-fluoromethyl-prednisone is obtained.

Example 10 (a) According to the method described in Example 1(a), the 6a-methyl-l6-fluoromethyl-15-dehydro-17a-hyof chloranil.

droxy-progesterone is obtained from 6a-methyl-16-fluoromethyl-16a,17a-oxido-progesterone.

(b) According to the methods described in Example 7(b) and (c), the 6a-rnethyl-l6-fluoromethyl-4,IS-pregnadiene-l7a,2l-diol-3,20-dione is obtained from 60cmethyl 16 fluoromethyl 15 dehydro 17a hydroxyprogesterone.

m. 240 u, l dm. 420

(c) According to the method described in Example 8(a), the 6ot-methyl-l fi-fluoromethyl-lS-dehydro-hydrocortisone is prepared from 6a-methyl-l6-fluoromethyl- 4,15-pregnadiene-17a,21-diol-3,20-dione.

(d) According to the method described in Example 7(h), the Got-methyl-16-fluoromethyl-IS-dehydro-prednisolone is prepared from Got-methyl-16-flu0romethyl-15-dehydro-hydrocortisone.

max. l"; i lm.

(e) According to the method described in Example 4(b), the 6a-methyl-1-fluoromethyl-15-dehydro-hydrocortisone-Zl-acetate (acetylated as described in Example 9(d)) is dehydrogenated to form 6a-methyl-16-fluoromethyl 4,6,15 pregnatriene-l1p,17a,21-triol-3,20-dione- 21-acetate.

(f) The product of Example 10(e) is dehydrogenated according to the method described in Example 8(e whereby the 6u-methy1-16-fluoromethyl-1,4,6,15-pregnatetraene-l1B,17a,21-triol-3,20-dione-2l-acetate is obtained.

The corresponding 21-alcohol is prepared by saponification of this product as described in Example 9(b).

Example 11 According to the methods set forth in Example 10, the *6e-methy1-l6-ch1oromethyl-1S-dehydro-prednisolone and the 6a-methyl-l6-chloromethyl-l,4,6,l5-pregnatetraene- 11,8,l7a,2l-triol-3,20-dione are obtained from 6a-methyl- 16fl-chloromethyl-l6u, 17a-oxido-progesterone.

Example 12 According to the methods set forth in Example 10, the 6e-fluoro-l6-fluoromethyl-IS-dehydro-prednisolone is obtained from 6a-fiuoro-l6 8-fluorornethyl-16a,l7 t-oxidoprogesterone.

max. l; izim. Example 13 According to the methods set forth in Example 10, the fia-fiuoro-l6-chloromethyl-1S-dhydro-prednisolone is obtained from 6a-fluoro-l6B-chloromethyl-16a,l7a-oxidoprogesterone.

max. l) lzz m. Example 14 (a) 16 chloromethyl-lS-dehydro-17a-hydroxy-progesterone is prepared in accordance with the method of Example 5. M.P. 198-199" C.

(b) The product obtained in Example 14(a.) is subjected to the action of Curvularia lunata according to the procedure set forth in Example 8(c) to form l6-chloromethyl-4, -pregnadiene-1 lB,17a-diol-3,20-dione.

max. y; tits.

(c) 16 chloromethyl 4,15-pregnadiene-l15,17a-dio1- 3,20-dione was treated in accordance with the method described in Example 7 (b) to form 16-chloromethyl-4,15-

. pregnadiene-l1B,17a,21-triol-3,20-dione-2l-acetate.

(d) The solution of 2 g. of 16-chloromethyl-4,lS-pregnadiene-l1B,l7a,2l-triol-3,20-dione-2l-acetate in 60 ml. of tetrahydrofuran is refluxed for 12 hours with 1.58 g. The reaction mixture is diluted with water and extracted with chloroform. .The combined chloro- A 282.5 ma, Elfi 690 (1) According to the procedure set forth in Example 8(e the 16 chloromethyl 4,6,15 pregnatriene- 11,8,17a,2l-triol 3,20-dione-2l-acetate is dehydrogenated to form 16 chloromethyl 1,4,6,15 pregnatetraene- 11,8,17a,21-triol-3,20-dione-2l-acetate.

(g) As described in Example 7(g), the 16-chlorornethyl 1,4,6,15 pregnatetraenel1B,17e,21-triol3,20-dione- 2l-acetate is saponified to prepare the l6-chloro-methyl- 1,4,6,'1 5-pregnatetraene-l 1,8,17a,2il-triol-3,20-dione.

max. l: izfm. Example 15 The procedure of Example 14 is applied identically to 1618-fluorometl1yl16 17a-oxidoprogesterone to prepare l6 fiu-oromethyl 4,6,15-pregnatriene-l1fi,17e,21-trio1-3, ZO-dione and 16-fluoromethyl-1,4,6,15-pregnatetraene- 1lfl,17a,2latriol-3,20-dione.

Example 16 (a) According to the methods given in Example 7(a) to 7(a), the 16 fiuoromethyl-ll-epi-15-dehydroahydrocortisOne-ZI aceta-te is prepared from lp-fluoromethyl- 1 6e,17a-oxido-progesterone.

(b) The 16 fluoromethyl-ll-epi-15-dehydro-hydrocortisone-Zl-acetate is dissolved in 25 ml. of chloroform and 25 ml. of pyridine. The solution is cooled to 0 C. and 7 g. of p-toluene-sulfonic acid are added with stirring. The mixture is stirred for another 2 hours at 0 C. and is allowed to stand overnight at room temperature. Then it is poured into water and extracted several times with chloroform. The chloroform extracts are neutralized and dried. Upon evaporation, the 16-iiuoromethyl-11-epiv 15dehydrohydrocortisonel l-tosylate-Zl-acetate is recrystallized from methanol.

The ester thus obtained is dissolved in ml. of glacial acetic acid and refluxed for 30 minutes with 9 g. of anhydrous sodium acetate. The mixture is poured into 500 ml. of water. The precipitated l6-fluoromethyl-4, 9 l 1 ,15 pregnatriene-l7ot,2l-diol-3 ,20-dione-21-acetate is filtered off and recrystallized from ethyl acetate.

(c) 7.8 g. of l6-fiuoromethyl4,9(1l),l5-tpregnatriene- :,21-di0l-3,20 clione-21aoetate are dissolved in 315 ml. of dioxane and 40 ml. of water. After addition of 4.55 g. of N-bromo-succinimi-de and 1.68 ml. of perchloric acid (70%), the mixture is allowed to stand for 1 hour at room temperature. Then it is poured into Water. The precipitated 16-fiuoromethyl-9abromo-1S-dehydrohydrocortisone-Zl-acetalte is filtered otf, Washed with Water and dried.

(d) The crude 16fiuoromethyl-9a-brorno-1S-dehydrohydrocortisone-Zl-acetate is dissolved in 45 0 ml. of ethanol and refluxed for 2 hours with 19 g. of potassium acetate. The mixture is poured into water and the obtained emulsion is extracted several times with chloroform. The combined chloroform extracts are worked up in the usual way. Upon evaporation, the 16-fluorornethyl-9p,l1/3-oxido-4, 15-pregnadiene-17a,2ldiol-3,20-dione-21-acetate is recrys- \tallized from methanol.-

(e) 4.2 g. of 16-fluorornethyl-9fl,l1 6-oxido-4,15-pregnadiene-170;,21-diol-3,2O-dione-21acetate are dissolved in 1 l 42 ml. of anhydrous chloroform and added at -60 to 25 ml. of a mixture of 40 ml. of tetrahydrofuran, 15 ml. of chloroform and 25 g. of hydrogen fluoride. The reaction mixture is allowed to stand for 4 hours at 30 and for 4 hours at The solution is poured into a solution of NaHCO The steroid is extracted with chloroform. Upon evaporation, the 9ec-fl1101'0l 6-fluoromethyl-15-dehydro-hydrocortisone-2l-acctate is recrystallized from acetone.

Am. 239 m li... 3

(1) 9a fluoro-16-fluoromethyl-IS-dehydro-hydrocortisone-acetate is saponified according to Example 7(g) to prepare 90c fiuoro-l6-fluoromethyl-lS-dehydro-hydrocortisone.

(g) According to the method described in Example 8(e the 900 flu-oro-l6-fiuorornethyl-1S-dehydro-hydrocortisone is dehydrogenated by the action of Bacillus splzaericus to form 9e-fiuoro-16-fiuoromethyl-lS-dehydroprednisolone.

A 239 m E93, 410 Example 17 According to the method described in Example 16, the 9ot-fluoro-16-chloromethyl-1S-dehydro-prednisolone is prepared from l6t3-chloromethyl-16a,17a-ox-ido-progesterone.

ruux. Example 18 Am... 240 u, li... 415

(c) g. of Got-methyl-16-fluoromethyl-4,15-pregnadiene-3,20-dione-17a-ol-acetate are refluxed for 7 hours with 100 ml. of methylethylketone and 7 g. of chloranil. Upon cooling, the solution is poured into water and extracted with chloroform. The chloroform extract is washed subsequently with Water, aqueous sodium hydroxide (1%) and again water and is dried with sodium sulfate. Upon evaporation, the 6 methyl 16 fluoromethyl 4.6,15 pregnatriene-17ot-ol-3,20-di0ne-l7-acetate is crystallized from methanol.

Example 19 (a) 8.5 g. of 16-fluorornethyl-4,IS-pregnadiene-l1(5', 17c,21-triol-3,ZO-diOne-ZI-acetate obtained according to Example 9(d) are refluxed for 7 hours with 335 ml. of tert.butanol and 59 g. of chloranil. The solution is concentrated to a volume of 100 m1. and extracted exhaustedvly with chloroform upon addition of 380 ml. of water.

The combined chloroform extracts are treated with ice cooled aqueous sodium hydroxide, washed with water to neutrality and concentrated under reduced pressure. The residue is dissolved in 25 ml. of a mixture containing equal volumes of benzene and of chloroform and chromatographed through 370 g. of fiorisil. The column is eluated with benzene/chloroform (1:1). The eluates 20 to 50, each of 200 ml., are combined and concentrated.

12 The 16-fiuoromethyl-4,6,IS-pregnatriene-I 1B,17a,21-trio'l- 3,20-dione-21-acetate is recrystallized from ethyl acetate.

(b) 5.8 g. of 16-fluoromethyl-4,6,lS-pregnatriene-l1/3, l7z,21-triol-3,20-dione-21acetate are dissolved in ml. of methanol and refluxed for 14 minutes upon addition of a hot solution of 1.28 g. of sodium bicarbonate in 19 ml. of water. The cooled solution is poured into a mixture of 900 ml. of water and 3 ml. of acetic acid, and extracted with chloroform. The combined extracts are washed with water and concentrated. The 16-fluoromethyl-4,6, 15-pregnatriene-11{3,17a,21-triol3,20-di0ne is recrystallized from methanol.

(c According to the method described in Example 7(11), 5 g. of 16-fiuoromethyl-4,6,l5-pregnatriene-1lfi, 17a,2l-triol-3,20-dione are dehydrogenated whereby the 16 fiuoromethyl-1,4,6,15-pregnatctraene-11[i,17a,21-triol- 3,20-dione is obtained.

A 220, 255, 298 mp, E 3, 330, 243, 324

(c;,) According to the method described in Example 2( 5 g. of 16-fluoromethyl-4,6,1S-pregnatriene-llfl, 17a,21-triol-3,20-dione are dehydrogenated within 22 hours. The obtained 16-fiuoromethyl-1,4,6,15-pregnatetraene-llfl,17a,21-triol-3,20-dione is recrystallized from ethyl acetate.

A... 220, 255, 298 m E1 2 330, 243, 324 Example 20 (a) 1.5 g. of 6-Cl1l0r0-lQB-fiUOIOmGthYl-l60,17ot-0Xld0- 4,6-pregnadiene-3,ZO-dione are dissolved in 23 ml. of acetone. The solution is allowed to stand for 2 hours at room temperature with 7.5 ml. of sulfuric acid (35%). The reaction mixture is worked up as described in Example 1 whereby the 6-ehlor0-16-fiuoromethyl-4,6,15-

. pregnatriene-3,20-dione-1'7cz-ol is obtained.

NW. 8 m ll... 5

(b) According to the method described in Example 18(1)), the 6-chloro-l6-fiuoromethyl-4,6,IS-pregnatrienel7u-Ol3, 20-Cli0t16 is acetylated to form 6-chloro-16-fluoromethyl-4,6,15-pregnatriene-17ot-ol-3,20-dione-1'7-acetate.

A 285 m Ei'itm. 550

The corresponding 16-chlorornethyl-compound is obtained when using 6-chloro-16fl-chloromethyl-16a,17aoxido-4,6-pregnadiene-3,20-dione as starting material.

Example 21 A variety of esters of the compounds prepared in accordance with the procedures of the preceding examples are prepared by treating the free alcohols with acylating agents by conventional methods. Thus, the hydroxyl group in the 21-position of the following compounds is esterified:

15 -dehydro- 1 6-fluoromethyl-prednisolone 1 S-dehydro- 1 6-chloromethyl-prednisolone 15-dehydro-16-fluoromethyl-prednisone a-methyl-lS-dehydro-l6-fiuoromethyl-prednisolone 6c4-methyl-15-dehydro-16-chloromethyl-prednisolone 6o;-fluoro-1 S-dehydro-16-fiuoromethyl-prednisolone 9a-fluoro-1S-dehydro-16-fiuoromethyl-prednisolone 6,1S-bis-dehydro-l6-fluoromethyl-prednisolone 6,IS-bis-dehydro-16-luoromethy1-hydrocortisone.

The esthers include the phosphate and the sodium salts thereof; the hemisulfate and the sodium salt thereof; the hemisuccinate and the sodium salt thereof; the diethylaminoacetate and the hydrochloride thereof; the acetate; the tert.butylacetate; the trimethylacetate; and the metasulfobenzoate.

Likewise, the Not-acetates and 17a-capronates of 15- dehydro-l6-fluoromethyl-17a-hydroxy progesterone, 60-

I O O Tenn:

and the A -derivatives thereof, wherein R is a member of the group consisting of hydrogen and the acid moiety of an aliphatic carboxylic acid containing up to 8 carbon atoms;

R is a member of the group consisting of OH and H;

R; is a member of the group consisting of hydrogen, methyl and chlorine, with the provision that when R represents OH, R is hydrogen; and

X is a member of the group consisting of chlorine and 2. A pharmaceutical composition containing from 0.1 to 70 mg. of a compound as claimed in claim 1 together with a pharmaceutically acceptable carrier.

3. 15 dehydro-16-fluoromethyl-17a-acetoxy-progesterone.

4. a methyl 16-fluoromethyl-4,15-pregnadiene-17aol-3,20-dione-17-acetate.

5. 16 fiuoromethyl 4,6,15 pregnatriene-17a-ol-3,20- dione-17-acetate.

6. 6 chloro 16-fluoromethyl-4,6,15-pregnatriene-17a- 01-3,20-dione-17-acetate.

References fiited by the Examiner UNITED STATES PATENTS 2,883,379 4/59 Moreland 260-239.55 3,057,884 10/62 Walker et a1. 260397.3 3,065,239 11/62 Wendler et al. 260397.45

LEWIS GO'ITS, Primary Examiner.

UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. 3,183,150 May 11, 1965 Fritz von Werder et al.

It is hereby certified that error appears in the above numbered patent requiring correction and that the said Letters Patent should read as corrected below.

Column 1, line 20, for "includes" read include line 43, for "aydrogen" read hydrogen same column 1, lines 50 to 62, the formula should appear as shown below instead of as in the patent:

column 3, line 3, for "optiumum" read optimum line 9, for "from" read form line 31, after "dioxane" strike out the comma; same column 3, line 46, for "fermentatioin" read fermentation column 6, line 63, for "epic-" read epiline 72, for "an" read and column 7, line 6, after "CrO insert a closing parenthesis; same line 6, for

"bleow 10 A." read below 10 C. same column 7, line 40, for "-oxide" read oxidocolumn 9, line 51, for "-dhydro-" read dehydrocolumn 10, lines 44 and 60, for "16-fiuoromethyl", each occurrence, read l6- fluoromethyl- Signed and sealed this 12th day of October 1965.

(SEAL) Attest:

ERNEST W. SWIDER EDWARD J. BRENNER Attesting Officer Commissioner of Patents 

1. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF
 2. A PHARMACEUTICAL COMPOSITION CONTAINING FROM 0.1 TO 70 MG. OF A COMPOUND AS CLAIMED IN CLAIM 1 TOGETHER WITH A PHARMACEUTICALLY ACCEPTABLE CARRIER. 